Protein vesicles and methods of making and using thereof

Abstract

Hollow, protein vesicles are provided. The protein vesicles can be loaded with desired cargo to be delivered to a subject, for example a human. One embodiment provides a protein vesicle made of a protein membrane surrounding a hollow core. The protein membrane includes a first and a second modular protein amphiphile. The first modular protein amphiphile includes a hydrophobic block and a first hydrophilic protein binding block. The second modular protein amphiphile includes a variable polypeptide block and a second hydrophilic protein binding block that binds to the first hydrophilic protein binding block. The first and second protein amphiphiles self-assemble to form the hollow, protein vesicle.

We claim: 
     
         1 . A hollow, protein vesicle comprising:
 a protein membrane surrounding a hollow core, wherein the protein membrane comprises a first and a second modular protein amphiphile, wherein the first modular protein amphiphile comprises a hydrophobic block and a first hydrophilic protein binding block and the second modular protein amphiphile comprises a variable polypeptide block and a second hydrophilic protein binding block that binds to the first hydrophilic protein binding block; and   wherein the first and second protein amphiphiles self-assemble to form the hollow, protein vesicle.   
     
     
         2 . The hollow, protein vesicle of  claim 1 , wherein the hydrophobic block of the first modular protein amphiphile comprises elastin-like polypeptide. 
     
     
         3 . The hollow, protein vesicle of  claim 2 , wherein the first and second hydrophilic protein binding blocks comprise a leucine zipper motif. 
     
     
         4 . The hollow, protein vesicle of  claim 3 , wherein the first hydrophilic protein binding block comprises an arginine-rich leucine zipper motif and the second hydrophilic protein binding block comprises a glutamic acid-rich leucine zipper motif. 
     
     
         5 . The hollow, protein vesicle of  claim 1 , wherein the second hydrophilic block comprises a catalytic domain of an enzyme, a targeting moiety, a fluorescent polypeptide, a Human Leukocyte Antigen polypeptide, a T cell receptor polypeptide, a detectable label, a globular polypeptide, an immunostimulatory polypeptide, or combinations thereof. 
     
     
         6 . The hollow, protein vesicle of  claim 5 , wherein the enzyme is selected from the group consisting of a dehydrogenase, a reductase, a protease, a synthetase. 
     
     
         7 . The hollow, protein vesicle of  claim 5 , wherein the targeting moiety is selected from the group consisting of an antibody, aptamer, ligand receptor, and fusion protein. 
     
     
         8 . The hollow, protein vesicle of  claim 7 , wherein the antibody is selected from the group consisting of a monoclonal antibody, a polyclonal antibody, a diabody, a single chain antibody, and antigen binding fragments thereof. 
     
     
         9 . A method for making hollow, protein vesicles comprising:
 combining a plurality of first and second modular protein amphiphiles in a solution on ice;   adjusting the salt concentration to promote or allow inverse phase transition of the protein amphiphiles; and   incubating the salt-adjusted solution at room temperature to form hollow, protein vesicles.   
     
     
         10 . The method of  claim 9 , wherein the first modular protein amphiphile comprises a hydrophobic block and a first hydrophilic protein binding block and the second modular protein amphiphile comprises a variable polypeptide block and a second hydrophilic protein binding block that binds to the first hydrophilic protein binding block. 
     
     
         11 . The method of  claim 10 , wherein the hydrophobic block of the first modular protein amphiphile comprises elastin-like polypeptide. 
     
     
         12 . The method of  claim 11 , wherein the first and second hydrophilic protein binding blocks comprise a leucine zipper motif. 
     
     
         13 . The method of  claim 12 , wherein the first hydrophilic protein binding block comprises an arginine-rich leucine zipper motif and the second hydrophilic protein binding block comprises a glutamic acid-rich leucine zipper motif. 
     
     
         14 . A hollow, protein vesicle produced according to the method of  claim 9 . 
     
     
         15 . A loaded, protein vesicle comprising:
 a protein membrane surrounding a core, wherein the protein membrane comprises a first and a second modular protein amphiphile, wherein the first modular protein amphiphile comprises a hydrophobic block and a first hydrophilic protein binding block and the second modular protein amphiphile comprises a variable polypeptide block and a second hydrophilic protein binding block that binds to the first hydrophilic protein binding block; and   wherein the first and second protein amphiphiles self-assemble around cargo to form the protein vesicle with the cargo loaded in the core of the protein vesicle.   
     
     
         16 . The loaded, protein vesicle of  claim 15 , wherein the hydrophobic block of the first modular protein amphiphile comprises elastin-like polypeptide. 
     
     
         17 . The loaded, protein vesicle of  claim 16 , wherein the first and second hydrophilic protein binding blocks comprise a leucine zipper motif. 
     
     
         18 . The loaded, protein vesicle of  claim 17 , wherein the first hydrophilic protein binding block comprises an arginine-rich leucine zipper motif and the second hydrophilic protein binding block comprises a glutamic acid-rich leucine zipper motif. 
     
     
         19 . The loaded, protein vesicle of  claim 16 , wherein the cargo comprises a drug, a protein, a contrast agent, nanoparticles, a fluorophore, a radiolabel, a therapeutic compound, an antioxidant, a growth factor, a cytokine, a chemoattractant, a nucleic acid or a combination thereof. 
     
     
         20 . The loaded, protein vesicle of  claim 19 , wherein the drug is selected from the group consisting of a cytocidal compound, a chemotherapy agent, a cellular metabolism blocker, a cell cycle inhibitor, or a combination thereof. 
     
     
         21 . The loaded, protein vesicle of  claim 19 , wherein the nucleic acid encodes a functional protein. 
     
     
         22 . The nucleic acid of  claim 19 , wherein the nucleic acid is selected from the group consisting of double stranded DNA, RNA, siRNA, microRNA, single stranded DNA, and anti-sense DNA.